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Journal of Sun Yat-sen University(Medical Sciences) ; (6): 467-472, 2019.
Article in Chinese | WPRIM | ID: wpr-817779

ABSTRACT

@#【Objective】To screen survival-related differential expression of long non-coding RNA(lncRNA)and its co-expressed genes in breast cancer patients and to verify their expression in breast cancer cells.【Methods】RNA-seq data of 943 cases(837 breast cancer + 106 normal controls)by the TCGA database were screened,and found that long non-coding MAPT-AS1 highly expressed,and breast cancer patients had longer survival. The long non-coding MAPT- AS1 overexpression and interference plasmid was constructed,and the constructed plasmid was transfected into breast cancer cell line T47D,and the stably expressed T47D cell line was screened by puromycin. The expression of long non-coding MAPT-AS1 and its co-expressed genes was verified by the methods of RT-qPCR.【Results】Fluorescence microscopy and RT-qPCR confirmed that the long non-coding MAPT-AS1 overexpression and interference-transfected breast cancer cell line T47D were successfully constructed,and the long non-coding MATS-AS1 interference fragment shRNA3 with the highest interference efficiency was screened. The expression of MAPT ,MAPT- IT1 and NXNL2 in the co-expressed gene was decreased after transfection of the shRNA3 interference fragment ,which was consistent with the expression trend of the long non-coding MAPT-AS1.【Conclusion】The long non-coding MAPT-AS1 overexpression and interference plasmid transfected breast cancer cell line T47D were successfully constructed,and the expression of the co- expressed gene was consistent with the database. The study laid the foundation for further study of the mechanism of action of long non-coding MAPT-AS1 gene in breast cancer.

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